Molecular Genetics of HSV Reactivation

HSV 再激活的分子遗传学

• 批准号: 6748099
• 负责人: David C. Bloom
• 金额: $ 26.66万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6748099
• 关键词: DNA binding protein; RNase protection assay; gene dosage; gene expression; genetic transcription; herpes simplex virus 1; laboratory rabbit; latent virus infection; methylation; nucleic acid sequence; pathologic process; polymerase chain reaction; regulatory gene; relapse /recurrence; virus genetics; virus replication

DESCRIPTION:(provided by applicant): Herpesviruses are ubiquitous and are responsible for significant human mortality and suffering both in terms of initial infections and (even more so) recurrences. In addition, with increasing occurrences of immunosuppressive disorders, a corresponding increase in the frequency of clinically significant initial and latent herpesvirus infections arise. Therefore, the long-term objective of this project is to gain a functional understanding of the herpesvinis-encoded genes involved in latency and reactivation - knowledge that is fundamental to the rational design of interventive therapies. We have identified a region of the genome termed the "reactivation critical region" (rcr) that is required for epinephrine-induced reactivation. The primary focus of this proposal is to characterize the functional role sequence elements in the rcr of the Herpes simplex virus type 1 (HSV- 1) genome play in reactivation of infections latent in sensory ganglia neurons. The overriding hypothesis of this study is that the HSV rcr facilitates reactivation by regulating gene expression from the latent genome allowing the initiation of acute gene expression during reactivation. Using a molecular genetic approach involving specifically engineered viral recombinants, we will follow the transcriptional and replicative processes of HSV- 1 reactivation in the rabbit cornea-epinephrine model to link viral genetics with processes in animals. Specifically, the proposed studies will test three potential mechanisms by which the rcr functions to facilitate reactivation: a) the rcr acts as a modulator of transcription which facilitates reactivation by regulating the expression of LAT, ICPO and/or ICP4, b) the rcr is a target of methylation and/or cellular factors that regulate transcription during latency and reactivation, and c) the rcr acts as a non-coding functional RNA that regulates gene expression via a dosage compensation-like mechanism. These three mechanisms are not mutually exclusive and it is likely an interplay between these regulatory elements is necessary for the maintenance of and reactivation from latency. This work will lead to the identification of viral target(s) of host factors that communicate stress stimuli leading to reactivation.

描述：（由申请人提供）：疱疹病毒无处不在，并且 造成重大人类死亡和痛苦的原因 初始感染和（更是如此）复发。此外，随着越来越多 免疫抑制疾病的发生，相应增加 有临床意义的初始和潜伏疱疹病毒感染的频率 出现。因此，该项目的长期目标是获得 对参与潜伏期的疱疹病毒编码基因的功能理解 和重新激活——合理设计的基础知识 介入治疗。我们已经确定了基因组的一个区域，称为 肾上腺素诱导所需的“重新激活关键区域”（rcr） 重新激活。该提案的主要重点是描述 1型单纯疱疹病毒rcr中的功能作用序列元件 (HSV-1) 基因组在感觉神经节潜伏感染的重新激活中发挥作用 神经元。 本研究最重要的假设是 HSV rcr 促进 通过调节潜在基因组的基因表达来重新激活，从而允许 重新激活期间急性基因表达的启动。使用分子 涉及专门设计的病毒重组体的遗传方法，我们将 跟踪 HSV-1 重新激活的转录和复制过程 兔角膜-肾上腺素模型将病毒遗传学与过程联系起来 动物。具体来说，拟议的研究将测试三种潜在的 rcr 发挥作用以促进重新激活的机制： a) rcr 作为转录调节剂，通过以下方式促进重新激活 调节 LAT、ICPO 和/或 ICP4 的表达，b) rcr 是 潜伏期调节转录的甲基化和/或细胞因子 和重新激活，以及 c) rcr 作为非编码功能性 RNA 通过类似剂量补偿的机制调节基因表达。这三个 机制并不是相互排斥的，它们之间可能存在相互作用 这些调节要素对于维持和重新激活是必要的 来自延迟。这项工作将导致病毒靶标的识别 传达应激刺激导致重新激活的宿主因素。